Screening method of indigenous group induction quenched bacteria and application thereof

A technology of quorum sensing and screening method, applied in bacteria and other directions, can solve the problems of affecting membrane separation performance, replacement frequency service life, increase MBR operating cost, and high membrane price, so as to reduce membrane fouling, simple and fast production, and prolong replacement cycle. Effect

Inactive Publication Date: 2018-11-23
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the problems of high membrane price and serious membrane pollution in the operation of MBR make its future development face severe challenges.
Among them, the problem of membrane fouling i...

Method used

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  • Screening method of indigenous group induction quenched bacteria and application thereof
  • Screening method of indigenous group induction quenched bacteria and application thereof
  • Screening method of indigenous group induction quenched bacteria and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] A screening method for quorum sensing quenching bacteria according to the present invention, such as Figure 4 As shown, the specific steps are as follows:

[0029] (1) From the activated sludge obtained from the reaction tank of the MBR device in the laboratory test, 1ml of the activated sludge was inoculated on the basic medium with a concentration of 2mM C6-AHL as the only carbon source, and after 5 days of enrichment culture Finally, take 1% of the sample by volume and transfer it to a new basic medium containing C6-AHL for cultivation, and repeat the above steps for 3 times;

[0030] (2) Pick an appropriate amount of cultured bacteria liquid and streak culture on LB solid medium, culture at 30°C for 10-14 hours, repeat the above steps 2-3 times to purify the bacteria, and pick a single colony for storage .

[0031] (3) The basic medium composition in the above steps is:

[0032] Major elements (g / L): NaCl 1.0, KCl 0.5, MgCl 2 ·6H 2 O 0.4, CaCl 2 2H 2 O 0.1, ...

Embodiment 2

[0038] The quorum induction quenching bacteria screened by the present invention is used to degrade the N-acyl homoserine lactone in the activated sludge, comprising the following steps:

[0039] Pick the quorum sensing quenched bacterial colony, inoculate it into LB liquid medium for overnight culture, and cultivate it at a constant temperature of 30°C for 10-14 hours, take the mixed solution and centrifuge it, wash the centrifuged bacteria repeatedly with normal saline, and then use ultrapure water to Bacteria are diluted to a bacterial solution with OD600=1.0;

[0040]Mix the bacterial solution with C6-AHL and C8-HSL at a ratio of 100:1, so that the final concentration of C6-AHL and C8-HSL in the mixed solution is 100ng / mL, and the mixed solution is placed in a constant temperature shaking incubator at 30°C Shake with the condition of 120rpm, and take samples at 10min, 20min, 60min and 120min time points respectively, measure the remaining C6-AHL and C8-HSL concentration in...

Embodiment 3

[0045] Pick colonies of the quorum quenching bacteria and inoculate them into LB liquid medium, culture them in a constant temperature incubator at a constant temperature of 30°C for 10-14 hours, take the mixed solution and centrifuge, wash the centrifuged bacteria repeatedly with physiological saline for 3 times, Discard the supernatant, weigh 0.1g of bacteria (wet weight), embed the bacteria with a PVDF flat film, seal the flat film with epoxy resin and place it in the air to solidify so that it becomes a dumpling-shaped bacterial bag, and the bacterial bag is formed later as image 3 shown.

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Abstract

The invention relates to a screening method of indigenous group induction quenched bacteria and a purpose, and belongs to the technical field of strain screening, wherein the group induction quenchedbacteria are screened by the following steps of screening out strains using N-acyl-homoserine lactone as a unique carbon source to grow from active sludge sampled from an indigenous MBR device; verifying the N-acyl-homoserine lactone degradation capability of the screened group induction quenched bacteria; embedding the group induction quenched bacteria by a PVDF flat plate film to prepare a bacterium bag; verifying the N-acyl-homoserine lactone degradation capability of the prepared group induction quenched bacterium bag. The product has a good degradation effect on the N-acyl-homoserine lactone. The product has the advantages that the used raw materials are safe and non-toxicity; the environmental-friendly effects are achieved; the manufacturing is simple, fast and convenient; the separation and recovery from the active sludge mixed solution are easy, and the like. The screening method can be used for the film pollution control in the MBR waste water treatment process and the like.

Description

technical field [0001] The present invention designs strain screening technical field, and specifically relates to a screening method and application of native quorum induction quenching bacteria. Background technique [0002] Membrane bioreactor (MBR) is a water treatment technology that combines membrane separation unit and biological treatment unit efficiently. The obvious advantage of convenience. However, the problems of high membrane price and serious membrane pollution in the operation of MBR make its future development face severe challenges. Among them, the problem of membrane fouling is particularly prominent. Membrane fouling will affect the separation performance, replacement frequency, and service life of the membrane, which will significantly increase the operating cost of the MBR. Soluble or colloidal substances produced by microbial metabolism are adsorbed on the surface of the membrane and in the voids. Bacteria adsorb on the surface of the membrane and p...

Claims

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Application Information

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IPC IPC(8): C12N1/20
CPCC12N1/20
Inventor 黄瑾辉顾岩岭曾光明史亚慧胡熠
Owner HUNAN UNIV
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